CELL COUNTING ISSUE
All-in-One Fluorescence Microscope BZ-X800E
ALL-IN-ONE FLUORESCENCE MICROSCOPE – NO DARKROOM REQUIRED
CONCEPT·FEATURES·APPLICATIONS
5VOL.
An easier approach to fluorescence microscopy
Advanced functionality without compromising simplicity
Image Quality:Observe high-resolution optical slices comparable to a confocal laser microscope.
Application Diversity:Combine the functionality of a slide scanner and time-lapse incubator on a single platform.
Analysis Capability:High-throughput quantification software for high-content data acquisition.
Install the microscope in a location that provides the
highest experiment efficiency.
All operations are performed in the software. Any user can easily capture images and reproduce
results.
High-sensitivity monochrome cooled CCD camera provides high resolution images
in fluorescence, brightfield, and phase contrast.
Built-in Darkroom Clear ImagesFull Electronic
Controls
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P.4 CARDIAC FIBROSISMeasuring the cardiac fibrosis ratio of a crab-eating macaque
COLONYP.5 Counting iPS cell colonies in a well plate
P.6 FAT CELLQuantitative analysis of adipocytes
P.8 STOMACH CANCERQuantification of immunostained stomach cancer tumor sections
P.7 FISHCounting chromosomes in a nucleus
P.9 CULTIVATED CELLSMeasuring the transfection efficiency of cultivated cells
TA B L E O F C O N T E N T S
All-in-One Fluorescence MicroscopeBZ-X800E
NEW
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BZ-X800E APPLICATION EXAMPLES
FIBROSISMeasuring the cardiac fibrosis ratio of a crab-eating macaque
Using the BZ-X800E
For a large section that cannot be seen within a single field-of-view, the motorized XY stage can capture and stitch together the entire region of interest in high resolution.
Fibrosis is a condition where tissues stiffen due to the deposition of collagen fibers.
As fibrosis progresses, normal organ function deteriorates.
IMAGE STITCHINGUp to 50,000 × 50,000 pixels can be joined together easily to capture a clear image
without stitch lines or brightness variations. Additionally, it is possible to accurately
quantify information such as fluorescent signal intensity.
Objective lens : CFI Plan Apo λ 4×Image stitching : 20 images × 12 images
AREA (TOTAL) AREA (FIBROTIC AREA) AREA PERCENTAGE
577,439,588 μm2 47,847,466 μm2 8.3%
COUNTEven for tilted specimens or specimens that have height differences, it is possible to create an image in which the entire specimen is in focus. This is accomplished by capturing multiple images in the Z-direction and stacking together only the parts that are in focus.
Hybrid Cell Count software can extract only fibrotic areas from the entire section and calculate the proportion automatically.
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BZ-X800E APPLICATION EXAMPLES
Objective lens:CFI Plan Apo λ 10× Image stitching: 7 images × 9 images
Batch measurement under the same conditions
COUNT 181
AVERAGE AREA 5,133 μm2
STANDARD AREA DEVIATION 8,730 μm2
TOTAL AREA 929,027 μm2
COUNT 251
AVERAGE AREA 8,315 μm2
STANDARD AREA DEVIATION 12,652 μm2
TOTAL AREA 2,086,967 μm2
COUNT 123
AVERAGE AREA 13,543 μm2
STANDARD AREA DEVIATION 17,023 μm2
TOTAL AREA 1,665,761 μm2
COUNT 171
AVERAGE AREA 12,969 μm2
STANDARD AREA DEVIATION 14,976 μm2
TOTAL AREA 2,217,727 μm2
COLONYCounting iPS cell colonies in a well plate
Induced pluripotent stem cells (iPS cells) are generated through the introduction of
specific genes into somatic cells. After cultivating for, they change into pluripotent
stem cells that have the ability to differentiate into cells of various tissues or organs and
proliferate almost without limit.
Using the BZ-X800E
For a well plate that cannot be seen within a single field-of-view,the motorized XY stage can capture and stitch together the entire region of interest in high resolution.
Hybrid Cell Count software allows extraction of only colonies from a well plate hole for automatic counting.
Multiple images can be processed collectively under the same conditions based on the features extracted with Hybrid Cell Count.
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BZ-X800E APPLICATION EXAMPLES
COUNT AVERAGE AREA STANDARD AREA DEVIATION
1485 169 μm2 403 μm2
Objective lens: CFI Plan Apo λ 20×
Batch measurement under the same conditions
COUNT
COUNT
1955
AVERAGE AREA
122 μm2
STANDARD AREA
DEVIATION
352 μm2
FAT CELLQuantitative analysis of adipocytes
Adipocytes are composed of droplets of fat called adipose tissue, and can be
classified into two types - unilocular (white) adipocytes and multilocular (brown)
adipocytes. A white adipocyte contains a single large fat droplet that stores fat.
A brown adipocyte contains multiple smaller droplets that burn fat to generate
heat. If white adipocytes grow in size due to an increase in obesity, they become
dysfunctional, and can lead to the onset of certain diseases.
Using the BZ-X800E
Each closely-spaced adipocyte can be extracted and counted using Hybrid Cell Count measurement software.
The image can be filtered by the area or shape of the extracted cells.
Multiple images can be processed collectively under the same conditions based on the features extracted with Hybrid Cell Count.
COUNT
3162
AVERAGE AREA
68 μm2
STANDARD AREA
DEVIATION
186 μm2
COUNT
1190
AVERAGE AREA
222 μm2
STANDARD AREA
DEVIATION
416 μm2
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BZ-X800E APPLICATION EXAMPLES
Signal G\R 0 1 2 3 4
0 40 5 1 1 0
1 23 35 13 1 0
2 10 24 18 2 2
3 0 5 7 2 0
4 1 3 4 2 1
Normal observation
COUNT
SECTIONING
No. of nuclei containing respective signals
Objective lens: CFI Plan Apo λ 100 × HSectioning + Z-stack
FISHCounting chromosomes in a nucleus
FISH (Fluorescence In Situ Hybridization) is a method of detecting the
distribution or amount of specific nucleic acids (DNA or RNA) using
fluorochromes. It is mainly used for detecting chromosomal abnormalities.
Using the BZ-X800E
The Optical Sectioning function makes it possible to eliminate fluorescence blurring and capture clear images.
A built-in Z-stack function captures multiple images at different focal positions and is able to create a fully-focused image by combining only the areas that are at their sharpest focus.
Hybrid Cell Count can be used to specify nuclei as mask areas to extract chromosomes contained in each nucleus and count them.
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BZ-X800E APPLICATION EXAMPLES
Objective lens: CFI Plan Apo λ 10×Image stitching: 20 images × 12 images
AREA (TOTAL) 65,426,415 μm2
AREA (TUMOR) 10,085,121 μm2
AREA PERCENTAGE 15.4%
COUNT
STOMACH CANCERQuantification of immunostained stomach cancer tumor sectionsImmunostaining, which is also called an immune antibody method or immunohistochemistry, is
often used for histopathological diagnosis. It binds a specific protein (antigen) to tissue with a
substance that specifically reacts to it (antibody) in order to visualize tumors or other target parts.
Using the BZ-X800E
For a large section that cannot be seen within a single field-of-view, the motorized XY stage can capture and stitch together the entire region of interest in high resolution.
Even for tilted specimens or specimens that have height differences, it is possible to create an image in which the entire specimen is in focus. This is accomplished by capturing multiple images in the Z-direction and stacking together only the parts that are in focus.
Hybrid Cell Count can extract only tumoral areas from the entire section and calculate the proportion automatically.
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BZ-X800E APPLICATION EXAMPLES
Counting cells using a phase contrast image
Counting expression events (fluorescent points) using cells as mask areas
Objective lens: CFI Plan Fluor DL 10× Cell 903
Expression 89
Efficiency 9.9%
CULTIVATED CELLSMeasurement of the transfection efficiency of cultivated cells
Transfection is a process of introducing nucleic acids into animal cells.
It is used to make cells introduce specific genes to express a protein of interest.
Using the BZ-X800E
It is possible to obtain phase contrast and fluorescence overlay images.
Even from a phase contrast image, our original measurement algorithm allows the outlines of cells to be extracted accurately.
Hybrid Cell Count can be used to extract the fluorescent protein contained in the cells and count them by using cells as mask areas.
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HYBRID CELL COUNTHigh-accuracy quantification
Mask functionBrightness (intensity) measurement
Extract only the sections with a brightness within a specific range to count the number or measure the area. Brightness variation correction and other functions ensure quantification of desired targets.
Color (hue) measurement
Extract only the sections with a color within a specific range to count the number or measure the area. Detailed and precise quantification is possible even with images with low contrast or clusters of cells.
Using the first specified area as a mask allows for example, counting the number of fluorescence-expressing cells or measuring the area of malignant tumor cells in normal tissue.
Phase contrast image (outline) measurement
The outlines of cells are accurately extracted even when the image has low contrast between the mea-surement target and background, as it the case with phase contrast images. Even adjacent cells can be counted accurately and easily.
Measurement results can be output in spreadsheet format as well as be displayed by the application software. This allows flexible data editing such as graph creation according to specific purposes.
Mask creation Measurement inside mask areas
Data can be output in spreadsheet format
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MACRO CELL COUNTReduce analysis time and improve data accuracy
QUANTIFYOBSERVE
Image 1
AUTO MEASUREMENT &DATA ACCUMULATION
Image 2 to ××
ANALYSIS
Image 1
QUANTIFYOBSERVE QUANTIFYOBSERVE
Image 2
QUANTIFYOBSERVE
Image 3
QUANTIFYOBSERVE
Image ××
DATA SUMMARIZATION QUANTIFY
Since each image is measured separately, conditions may differ.
REDUCED TIME
MORE RELIABLE MEASUREMENTS IN LESS TIMEOUTPUT CONDITIONS• Threshold value• Correction values• Mask setting• Measurement target range (upper/lower limits)• Colocalization setting, etc
All images are measured under the same conditions, eliminating user bias.
DRAG & DROP
CONDITIONFILE
CONVENTIONAL METHOD
MACRO CELL COUNT
SAVE CONDITION
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KA12-1017The information in this publication is based on KEYENCE’s internal research/evaluation at the time of release and is subject to change without notice. Company and product names mentioned in this catalog are either trademarks or registered trademarks of their respective companies.The specifications are expressed in metric units. The English units have been converted from the original metric units.
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